Applied Research -Summary / Abstract This proposal seeks continued funding for the Tetrahymena Stock Center to build the research capacity of the community of scientists that use this resource, by designing custom research materials for use in genetic studies. These activities center on two specific aims: 1) Promote future studies with Tetrahymena by developing and displaying new gene expression and mutation datasets on Stock Center informational resources. Our rationale for this approach is two-fold: first, the rapidly decreasing price of DNA sequencing technology makes new data-driven approaches very affordable; and second, recent improvements to Tetrahymena Genome Database (TGD) (such as the upgrade of the genome browser to JBrowse2) make it possible to offer analysis assistance to Stock Center users. Furthermore, because our resources are widely used in undergraduate classrooms to engage students in guided research experiences, we are obligated to offer resources that allow the next generation to train in data sciences. We will identify existing useful gene expression datasets deposited in NCBI databases for curation and visualization on the TGD genome browser. In addition, we will generate RNA-Seq data from key life cycle stages or growth conditions to fill in missing information. We will make these data accessible at TGD and on the WashU epigenome browser, a cutting- edge, genomic data visualization resource. In addition to gene expression data, we will use sequencing technologies to identify genes mutated in a selection of mutant strains with striking phenotypes. This will allow us to improve our resource curation by linking gene to phenotype with the hopes of stimulating new research. 2) Optimize gene knockout strategies for the strain engineering services offered by the Stock Center. As part of our function as a full-service resource center, the Stock Center creates a variety of genetic mutant strains for members of the research community. We will generate standardized workflows with predictable outcomes and time lines for the completion of custom gene knockouts in Tetrahymena. This will allow us to communicate more transparently and effectively with the patrons of this service. Overall, these enhancements to resources and practices will have the key outcome of ensuring that the Tetrahymena Stock Center can meet the research needs of both established and young investigators as they strive to make significant discoveries using ciliates.